Trans-acting transposase mutant from Tn5

Alison Delong, Michael Syvanen

Research output: Contribution to journalArticle

17 Scopus citations

Abstract

Transposition of Tn5 and of its component insertion sequence IS50R is regulated through the action of two proteins it encodes: a cis-acting transposase, Tnp, and a trans-acting inhibitor of transposition, Inh. The mechanism of the cis-acting Tnp and the relevance of inhibition to cis action have been addressed in the current study. A specific colony morphology assay for transposition of Tn5 was shown to be sensitive to Inh produced in trans and was used to screen for mutants in Inh and/or Tnp with altered regulation. A dominant mutant in IS50R that promotes transposition in trans was isolated and characterized. The mutant (449F) carries a Leu → Phe mutation at position 449 in Tnp. This mutation reduces the frequency of Tn5 or IS50R transposition in cis but allows Tnp-449F to act as efficiently in trans as it does in cis. Tnp-449F is sensitive to inhibition and, furthermore, Inh-449F is a competent inhibitor in trans. These results show that Tnp-449F is a trans-acting transposase, unlike wild-type Tnp, which is cis-acting.

Original languageEnglish (US)
Pages (from-to)6072-6076
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume88
Issue number14
StatePublished - Jul 15 1991

ASJC Scopus subject areas

  • General
  • Genetics

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